Detection of Mycoplasma in avian live virus vaccines by polymerase chain reaction.
The polymerase chain reaction (PCR) was evaluated to detect mycoplasma contamination of avian live virus vaccines. The specificity of the primers showed that 34 strains belonging to nine species of avian mycoplasma DNA could be detected. The sensitivity of PCR to detect mycoplasma DNA was 10(0.2) colony forming units (cfu) of Mycoplasma synoviae and 10(0.7) cfu of Mycoplasma gallisepticum. When M. synoviae and M. gallisepticum were spiked into several avian live virus vaccines, PCR gave a positive reaction except for the avian pox and the avian encephalomyelitis vaccines which were prepared from organ homogenates. Short-term incubation of avian encephalomyelitis vaccines improved the sensitivity of PCR to detect both M. synoviae and M. gallisepticum. Therefore, PCR, combined with the short-term incubation, were shown to be most effective in detecting mycoplasma contamination in all of avian live virus vaccines.
Kojima
A
A
National Veterinary Assay Laboratory, Tokyo, Japan.
Takahashi
T
T
Kijima
M
M
Ogikubo
Y
Y
Nishimura
M
M
Nishimura
S
S
Harasawa
R
R
Tamura
Y
Y
D000818:Animals; D001483:Base Sequence; D001717:Birds; D017931:DNA Primers; D004269:DNA, Bacterial / Q000302:isolation & purification; D004340:Drug Contamination; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014613:Vaccines, Attenuated; D014765:Viral Vaccines
D017931:DNA Primers; D004269:DNA, Bacterial; D014613:Vaccines, Attenuated; D014765:Viral Vaccines